Figure 4.
Platelet adhesion onto VWF under flow is reduced in the absence of α4A- and β1-tubulins. (A-B) Representative differential interference contrast (DIC) micrographs of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets rolling onto captured VWF at (A) 500 and (B) 3000 reciprocal seconds (s-1). Hirudinated whole blood was perfused into polydimethylsiloxane channels coated with the VWF-capturing peptide (vA3-III-23, 100 μg/mL). Blood was allowed to flow for 15 seconds before recording videos. For each micrograph, the dotted white circle indicates the initial position of 1 platelet, and the full white circle indicates its position after 800 ms. Scale bar: 10 μm. (C-D) Rolling velocity of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets onto captured VWF at (C) 500 and (D) 3000 reciprocal seconds (s-1). Velocities are expressed as μm/s and were determined based on DIC recordings. Bar graphs represent the mean ±SEM. Each symbol indicates an individual flow. N = 3 per strain, with ≥15 platelets counted per individual flow. Statistical analysis was conducted by 1-way ANOVA followed by Tukey HSD post-hoc test. (E-F) Platelet detachment from the VWF surface at (E) 500 and (F) 3000 reciprocal seconds (s-1) was determined by tracking individual KOB1 (B1) and DKO platelets during 30 seconds on DIC recordings. Results are expressed in %. Bar graphs represent the mean ±SEM. Each symbol indicates an individual flow. N = 3 per strain, with ≥15 platelets counted per individual flow. Statistical analysis was conducted by unpaired 2-tailed t test.

Platelet adhesion onto VWF under flow is reduced in the absence of α4A- and β1-tubulins. (A-B) Representative differential interference contrast (DIC) micrographs of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets rolling onto captured VWF at (A) 500 and (B) 3000 reciprocal seconds (s-1). Hirudinated whole blood was perfused into polydimethylsiloxane channels coated with the VWF-capturing peptide (vA3-III-23, 100 μg/mL). Blood was allowed to flow for 15 seconds before recording videos. For each micrograph, the dotted white circle indicates the initial position of 1 platelet, and the full white circle indicates its position after 800 ms. Scale bar: 10 μm. (C-D) Rolling velocity of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets onto captured VWF at (C) 500 and (D) 3000 reciprocal seconds (s-1). Velocities are expressed as μm/s and were determined based on DIC recordings. Bar graphs represent the mean ±SEM. Each symbol indicates an individual flow. N = 3 per strain, with ≥15 platelets counted per individual flow. Statistical analysis was conducted by 1-way ANOVA followed by Tukey HSD post-hoc test. (E-F) Platelet detachment from the VWF surface at (E) 500 and (F) 3000 reciprocal seconds (s-1) was determined by tracking individual KOB1 (B1) and DKO platelets during 30 seconds on DIC recordings. Results are expressed in %. Bar graphs represent the mean ±SEM. Each symbol indicates an individual flow. N = 3 per strain, with ≥15 platelets counted per individual flow. Statistical analysis was conducted by unpaired 2-tailed t test.

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